DNA-binding protein A (dbpA) has been reported associated with the pathogenesis and development of various cancers. However, no evidence showed the gene expression profiling alternation involved in dbpA knockdown in colorectal cancer (CRC) cells. Small interference RNA (siRNA) method was used to knock down dbpA expression in SW620 cells. The changes of gene expression profiles in dbpA knockdown SW620 cells were determined by microarray analysis and Western blot. A total of 578 genes expressed differentially (twofold change), 181 genes were up-regulated, and 397 down-regulated in the dbpA knockdown group in comparison with the control group. The discrimination reliability was further verified by principal component analysis and Pearson correlation. Gene ontology (GO) pathway analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis showed that the most significantly expressed genes were linked to MAPK signaling pathway. Furthermore, Western blot verified that dbpA knockdown directly inhibited the activations of TAK1, p38, and JNK in CRC cells. In conclusion, dbpA knockdown in SW620 cells altered the expression of carcinogenesis-associated genes in CRC and the involvement of dbpA on CRC might through MAPK signaling pathway, which might provide valuable evidence to further investigate the correlation between dbpA expression and CRC development.